RESEARCH OPPORTUNITY AWARD - CHARACTERIZATION OF NAG EXPRESSION AND FUNCTIONS

研究机会奖 - NAG 表达和功能的表征

• 批准号: 8167527
• 负责人: CARLA J GUTHRIDGE
• 金额: $ 1.39万
• 依托单位: UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8167527
• 关键词: Anthrax Vaccines; Anthrax disease; Antibodies; Antigens; Award; Bacillus anthracis; Bacillus anthracis spore; Binding; Biological Assay; Chimeric Proteins; Computer Retrieval of Information on Scientific Projects Database; Environment; Epitopes; Funding; Goals; Grant; Immune response; Immunity; Immunotherapeutic agent; In Vitro; Individual; Institution; Laboratories; Licensing; Maps; Monoclonal Antibodies; Morbidity - disease rate; Peptide Hydrolases; Peptides; Protein Binding; Recombinant Fusion Proteins; Research; Research Personnel; Resources; Serious Adverse Event; Source; Toxin; United States National Institutes of Health; Vaccination; Vaccines; Work; human monoclonal antibodies; improved; in vivo; mortality; three dimensional structure; vaccination strategy

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Bacillus anthracis spores can cause significant morbidity and mortality if released into the environment. There is a need for both an effective vaccine as well as targeted immunotherapeutic agents. The currently licensed vaccine requires six vaccinations, annual boosters, and has been associated with serious adverse events. Thus there has been a major effort made to develop an improved vaccination strategy which can generate lasting protective immunity with reduced vaccinations. Dr. James' laboratory has been working to identify the major humoral targets of B. anthracis that provide protection. Human monoclonal antibodies were previously generated from individuals who received the anthrax vaccine. Each of these monoclonal antibodies has previously been characterized in both in vitro and in vivo toxin neutralization assays as well as by linear epitope and protease fragment mapping studies. Multiple functionally informative anti-PA monoclonal antibodies were identified. However, the structural epitopes recognized by these functionally informative monoclonal antibodies remained unclear. These monoclonal antibodies apparently recognized conformational epitopes rather than the linear peptide epitopes of protective antigen. The goal of this study was to define the protective antigen structural determinants that were specifically recognized by the functionally informative monoclonal anti-PA antibodies. Using the three dimensional structure of anthrax PA as a guide, different domains of PA will be cloned as recombinant fusion proteins. These PA subdomain containing fusion proteins will then be expressed and used in protein binding assays to define which PA domains are required for anti-PA monoclonal antibody binding. The results of these studies will identify which domains of PA are the best targets for generating protective immune responses through vaccination.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 如果炭疽杆菌孢子释放到环境中，会导致严重的发病率和死亡率。需要有效的疫苗和靶向免疫治疗剂。目前获得许可的疫苗需要接种六次疫苗，每年加强接种，并且与严重的不良事件有关。因此，人们付出了巨大的努力来开发一种改进的疫苗接种策略，该策略可以通过减少疫苗接种来产生持久的保护性免疫力。詹姆斯博士的实验室一直致力于确定炭疽杆菌提供保护的主要体液靶点。人类单克隆抗体之前是从接受炭疽疫苗的个体中产生的。这些单克隆抗体中的每一种先前都已在体外和体内毒素中和测定以及线性表位和蛋白酶片段作图研究中进行了表征。鉴定出多种功能丰富的抗 PA 单克隆抗体。然而，这些功能信息丰富的单克隆抗体识别的结构表位仍不清楚。这些单克隆抗体显然识别构象表位而不是保护性抗原的线性肽表位。 本研究的目的是确定功能信息丰富的单克隆抗 PA 抗体特异性识别的保护性抗原结构决定簇。以炭疽PA的三维结构为指导，将PA的不同结构域克隆为重组融合蛋白。然后，这些含有融合蛋白的 PA 子结构域将被表达并用于蛋白结合测定，以确定抗 PA 单克隆抗体结合所需的 PA 结构域。这些研究的结果将确定 PA 的哪些域是通过疫苗接种产生保护性免疫反应的最佳靶标。